GlowCas9
Context:
Researchers from the Bose Institute, Kolkata, have developed a new engineered CRISPR protein called “GlowCas9”
This innovation allows scientists to observe the gene-editing process in real-time, addressing a long-standing challenge in the field.
About GlowCas9:
It is a bioluminescent version of the Cas9 protein (the molecular scissors used in CRISPR) that emits light while performing gene editing.
It was created by a team led by Dr. Basudeb Maji
It involves fusing the Cas9 protein with a split nano-luciferase enzyme derived from deep-sea shrimp proteins.
How does it Works?
The enzyme pieces attached to Cas9 remain inactive until the protein finds its target.
When Cas9 binds to the specific DNA sequence and folds correctly to cut it, the split enzyme pieces reconnect, restoring enzymatic activity and producing a visible light signal (similar to fireflies).
Significance & Applications:
Real-Time Tracking:
Unlike traditional methods that require destroying cells to verify editing, GlowCas9 allows scientists to visualize the process in living cells, tissues, and plant leaves as it happens.
Thermostability:
It is more stable than conventional enzymes and maintains activity at higher temperatures, which is crucial for successful gene therapy delivery.
Precision Medicine:
It enhances the precision of Homology-Directed Repair (HDR), a DNA repair mechanism vital for fixing hereditary mutations linked to diseases like sickle cell anemia and muscular dystrophy.
To demonstrate its precision, researchers used GlowCas9 to insert the DNA sequence corresponding to ACHARYA into a genome, honoring the institute's founder, Acharya Jagadish Chandra Bose.